Journal: The Journal of Biological Chemistry

Article Title: Pyroglutamate and O -Linked Glycan Determine Functional Production of Anti-IL17A and Anti-IL22 Peptide-Antibody Bispecific Genetic Fusions

doi: 10.1074/jbc.M112.417717

Figure Lengend Snippet: The peptide-antibody genetic fusion neutralizes the IL22 activity equivalent to parental anti-IL22 antibody while lacking anti-IL17A neutralizing activity. A, kinetic analysis with human IL22 using a Biacore 2000. B, IL22-stimulated release of GRO-α production in HT29 cells. C, table summary of kinetic analysis with human IL17A using a Biacore 2000. D, IL17A-stimulated release of GRO-α in BJ human foreskin fibroblast cells. Error bars, S.D.

Article Snippet: Bound GRO-α was detected with anti-human GRO-α biotin (catalog no. BAF275, R&D Systems) at 40 ng/ml, followed by streptavidin-HRP (catalog no. DY998, R&D Systems), followed by a 1:1 mixture of H 2 O 2 and tetramethylbenzidine substrate (catalog no. DY999, R&D Systems), followed by 2 n H 2 SO 4 .

Techniques: Activity Assay

Journal: BMB Reports

Article Title: Regorafenib prevents the development of emphysema in a murine elastase model

doi: 10.5483/BMBRep.2023-0072

Figure Lengend Snippet: Effect of regorafenib on PPE-induced in inflammatory mediators in BAL fluid. (A) Representative membrane images of the antibody array. (B) Quantification of relative optical density for IL-1β, IL-6, CXCL1/KC, and TIMP-1 in the array membranes. (C-F) Measurement of IL-1β (C), IL-6 (D), CXCL1/KC (E), and TIMP-1 (F) levels in BAL fluid by ELISA. Error bars indicate the mean ± SEM. N = 8 per group, *P < 0.05, **P < 0.01, ****P < 0.0001, t -test.

Article Snippet: Anti-IL-6 antibody (ab208113) from abcam (Cambridge, UK), CXCL1 antibody (12335-1-AP) from Proteintech (Rosemont, IL, USA), and TIMP-1 antibody (#AF980) from R&D Systems were purchased respectively.

Techniques: Membrane, Ab Array, Enzyme-linked Immunosorbent Assay

Journal: BMB Reports

Article Title: Regorafenib prevents the development of emphysema in a murine elastase model

doi: 10.5483/BMBRep.2023-0072

Figure Lengend Snippet: Effect of regorafenib on PPE-induced expression of inflammatory mediators in lung parenchyme. (A) Experimental scheme. Mice were sensitized by treatment with regorafenib (5 mg/kg) for one day in the only PPE + Reg group. After PPE treatment, regorafenib was administered orally to mice for six days. On the seventh day, lung tissues were removed from mice, prepared in paraffin sections and stained with antibodies for inflammatory mediators. (B) Representative images of stained paraffin sections. Red arrows indicate the macrophages in inserted boxes. Scale bars indicate 50 μm. (C-E) Quantification of . IL-6 (C), CXCL1/KC (D), and TIMP-1 (E). Error bars indicate the mean ± SEM. N = 5 per group, *P < 0.05, **P < 0.01, t -test.

Article Snippet: Anti-IL-6 antibody (ab208113) from abcam (Cambridge, UK), CXCL1 antibody (12335-1-AP) from Proteintech (Rosemont, IL, USA), and TIMP-1 antibody (#AF980) from R&D Systems were purchased respectively.

Techniques: Expressing, Staining

Journal: Oncology Letters

Article Title: CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer

doi: 10.3892/ol.2015.3614

Figure Lengend Snippet: Expression of chemokine (C-X-C motif) ligand in gastric cancer and para-carcinoma tissues, as determined by immunohistochemistry. (A) CXCL1-positive and (B) CXCL1-negative tumor tissue; and (C) CXCL1-negative normal tissue (streptavidin-peroxidase method; magnification, x200).

Article Snippet: Mouse anti-human CXCL1 monoclonal antibody (dilution, 1:50; catalog no., MAB275; R&D Systems, Inc., Minneapolis, MN, USA) and rabbit anti-human Snail monoclonal antibody (dilution, 1:50; catalog no., ab180714; Abcam, Cambridge, UK) were used.

Techniques: Expressing, Immunohistochemistry

Journal: Oncology Letters

Article Title: CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer

doi: 10.3892/ol.2015.3614

Figure Lengend Snippet: Univariate analysis of factors associated with the overall survival rate.

Article Snippet: Mouse anti-human CXCL1 monoclonal antibody (dilution, 1:50; catalog no., MAB275; R&D Systems, Inc., Minneapolis, MN, USA) and rabbit anti-human Snail monoclonal antibody (dilution, 1:50; catalog no., ab180714; Abcam, Cambridge, UK) were used.

Techniques: Expressing

Journal: Oncology Letters

Article Title: CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer

doi: 10.3892/ol.2015.3614

Figure Lengend Snippet: Multivariate analysis of CXCL1 expression associated with the overall survival rate.

Article Snippet: Mouse anti-human CXCL1 monoclonal antibody (dilution, 1:50; catalog no., MAB275; R&D Systems, Inc., Minneapolis, MN, USA) and rabbit anti-human Snail monoclonal antibody (dilution, 1:50; catalog no., ab180714; Abcam, Cambridge, UK) were used.

Techniques: Expressing

Journal: Oncology Letters

Article Title: CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer

doi: 10.3892/ol.2015.3614

Figure Lengend Snippet: Kaplan-Meier analysis of the overall survival time of patients with gastric cancer, based on CXCL1 and Snail expression. (A) High CXCL1 expression in gastric cancer tissue was associated with a poor overall survival time (P<0.001). (B) High Snail expression was also associated with a worse overall survival time (P<0.001). CXCL1, chemokine (C-X-C motif) ligand.

Article Snippet: Mouse anti-human CXCL1 monoclonal antibody (dilution, 1:50; catalog no., MAB275; R&D Systems, Inc., Minneapolis, MN, USA) and rabbit anti-human Snail monoclonal antibody (dilution, 1:50; catalog no., ab180714; Abcam, Cambridge, UK) were used.

Techniques: Expressing

Journal: Oncology Letters

Article Title: CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer

doi: 10.3892/ol.2015.3614

Figure Lengend Snippet: Association between CXCL1 and Snail expression levels and clinical characteristics of patients with gastric cancer.

Article Snippet: Mouse anti-human CXCL1 monoclonal antibody (dilution, 1:50; catalog no., MAB275; R&D Systems, Inc., Minneapolis, MN, USA) and rabbit anti-human Snail monoclonal antibody (dilution, 1:50; catalog no., ab180714; Abcam, Cambridge, UK) were used.

Techniques: Expressing

Journal: Oncology Letters

Article Title: CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer

doi: 10.3892/ol.2015.3614

Figure Lengend Snippet: Association between Snail expression and CXCL1 expression, determined using Fisher's exact test. CXCL1, chemokine (C-X-C motif) ligand.

Article Snippet: Mouse anti-human CXCL1 monoclonal antibody (dilution, 1:50; catalog no., MAB275; R&D Systems, Inc., Minneapolis, MN, USA) and rabbit anti-human Snail monoclonal antibody (dilution, 1:50; catalog no., ab180714; Abcam, Cambridge, UK) were used.

Techniques: Expressing

Journal: Oncology Letters

Article Title: CXCL1 expression is correlated with Snail expression and affects the prognosis of patients with gastric cancer

doi: 10.3892/ol.2015.3614

Figure Lengend Snippet: Multivariate analysis of CXCL1 and Snail expression associated with the overall survival rate.

Article Snippet: Mouse anti-human CXCL1 monoclonal antibody (dilution, 1:50; catalog no., MAB275; R&D Systems, Inc., Minneapolis, MN, USA) and rabbit anti-human Snail monoclonal antibody (dilution, 1:50; catalog no., ab180714; Abcam, Cambridge, UK) were used.

Techniques: Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Breaking the symmetry of cell contractility drives tubulogenesis via CXCL1 polarization.

doi: 10.1073/pnas.2315894121

Figure Lengend Snippet: Fig. 3. Soft matrix induces CXCL1 polarization promoting cell protrusions and tubulogenesis. Analysis of (A) Cxcl1 mRNA expression of cells grown on the fish skin–derived collagen gel–coating or gel for 6 h and their (B) CXCL1 protein level in the culture supernatant. **P < 0.01; ***P < 0.001. (C) Representative confocal microscopy images showing CXCL1 (green), pT18S19MLC2 (red), microtubules (cyan), and nucleus (blue) in cells on the fish skin–derived collagen gel or gel-coating for 6 h. (D) Statistical analysis of mean fluorescence intensity of CXCL1 per high power field (HPF) in C. There were approximately 61 and 104 cells per HPF in the gel-coating and gel group, respectively. ***P < 0.001. Analysis of (E) Cxcl1 mRNA expression of cells grown on the 9-mo-old rat tail collagen gel with or without sonication (75% amplitude) for 1 d and their (F) CXCL1 protein level in the culture supernatant. ***P < 0.001. (G) Representative epifluorescent microscopy images of cellular staining for microtubules (green), microfilaments (red), and nucleus (blue) in cells treated with SB225002 on the fish skin–derived collagen gel for 7 h (yellow arrows: cell protrusions). (H) Montage images extracted from timelapse recordings of cells treated with siRNAs on fish skin–derived collagen gel (Movie S4) (red arrows; cell protrusions at 6 h). (I) Statistical analysis of relative cell protrusion activity per high power field (HPF) in Fig. 4H. There were approximately 310, 267, and 247 cells per HPF at 1 h in the siCntl, siCxcl1, and siCxcr1 group, respectively. **P < 0.01. (J) Tube formation on fish skin–derived collagen gel for 2 d in cells with siRNA knockdown of CXCL1. Cells with siCntl served as control. ***P < 0.001. (K) Tube formation on fish skin–derived collagen gel for 1 d in cells treated with SB225002 CXCRs inhibitor. *P < 0.05; **P < 0.01. (L) Tube formation on the fish skin–derived collagen gel for 2 d in cells with siRNA knockdown of CXCR1. ***P < 0.001.

Article Snippet: Immunofluorescent staining was performed using specific antibodies against CXCL1 (AF- 515- NA, R&D Systems), tubulin- α (05- 829, Merck), pMLC2 (95777, Cell Signaling Technology), and E- cadherin (610182, BD Biosciences) followed by Alexa Fluor- conjugated secondary antibodies (Invitrogen).

Techniques: Expressing, Derivative Assay, Confocal Microscopy, Fluorescence, Sonication, Microscopy, Staining, Activity Assay, Knockdown, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Breaking the symmetry of cell contractility drives tubulogenesis via CXCL1 polarization.

doi: 10.1073/pnas.2315894121

Figure Lengend Snippet: Fig. 4. Reduction of cell contractility promotes cell protrusions. (A) Real-time PCR analysis of Cxcl1 expression in cells treated with or without blebbistatin on fish skin–derived collagen gel for 1 d. (B) ELISA of CXCL1 protein expression in the cell culture supernatant from cells treated with or without blebbistatin on fish skin–derived collagen gel for 1 d. (C) Representative confocal microscopy images showing CXCL1 (green), microtubules (red), microfilaments (cyan), and nucleus (blue) in cells treated with or without blebbistatin and grown on fish skin–derived collagen gel for 6 h. (D) Representative epifluorescent microscopy images of CXCL1 (green), microtubules (red), and nucleus (blue) in cells treated with or without blebbistatin on fish skin–derived collagen gel for 6 h. Cell morphology was observed through the bright field channel (Ph) (yellow arrows: cell protrusions). (E) Statistical analysis of cell protrusions per high-power field (HPF) of D. ***P < 0.001.

Article Snippet: Immunofluorescent staining was performed using specific antibodies against CXCL1 (AF- 515- NA, R&D Systems), tubulin- α (05- 829, Merck), pMLC2 (95777, Cell Signaling Technology), and E- cadherin (610182, BD Biosciences) followed by Alexa Fluor- conjugated secondary antibodies (Invitrogen).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Confocal Microscopy, Microscopy

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Breaking the symmetry of cell contractility drives tubulogenesis via CXCL1 polarization.

doi: 10.1073/pnas.2315894121

Figure Lengend Snippet: Fig. 6. Suppressing the mechanosensing machinery CD29 abolishes CXCL1 expression and tubulogenesis. (A) Western blot analysis showing the expression of CD29 and tubulin-α in cells transfected with control (shCntl) or CD29-targeting shRNA (shCD29). (B) Tube formation on fish skin–derived collagen gel for 1 d in cells with shCntl or shCD29. ***P < 0.001. (C) Representative confocal microscopy images of cells with phalloidin (red) for microfilaments and DAPI (blue) for the nucleus on FITC-conjugated collagen gel for 1 d. (D) Analysis of Cxcl1 mRNA expression in cells grown on fish skin–derived collagen gel for 1 d. ***P < 0.001. (E) ELISA analysis of cell culture supernatant taken from cells grown on fish skin–derived collagen gel for 1 d. ***P < 0.001. (F) Flow chart shows breaking the symmetry of cell contractility by reducing matrix stiffness or myosin activity promotes CXCL1 polarization at cell protrusions. CXCL1–CXCR1 interaction at cell protrusions promotes cell polarization and ECM remodeling, which in turn, accelerates cell–cell connection and tube elongation.

Article Snippet: Immunofluorescent staining was performed using specific antibodies against CXCL1 (AF- 515- NA, R&D Systems), tubulin- α (05- 829, Merck), pMLC2 (95777, Cell Signaling Technology), and E- cadherin (610182, BD Biosciences) followed by Alexa Fluor- conjugated secondary antibodies (Invitrogen).

Techniques: Expressing, Western Blot, Transfection, Control, shRNA, Derivative Assay, Confocal Microscopy, Enzyme-linked Immunosorbent Assay, Cell Culture, Activity Assay